QIAGEN's real-time PCR cycler, the Rotor-Gene Q, combines multiple optimized design features to provide the outstanding performance and reliable results that your research demands. Together with optimized QIAGEN kits for real-time PCR, the Rotor-Gene Q enables streamlined analysis for a wide range of applications. Q-Rex Software is the new operating and analysis software for life science qPCR applications, with a plug-in concept that lets you add new functionality without affecting established workflows.

The Rotor-Gene Q, if used in combination with QIAGEN kits indicated for use with the Rotor-Gene Q instrument, is intended for the applications described in the respective QIAGEN kit handbooks. If the Rotor-Gene Q instrument is used with kits other than QIAGEN kits, it is the user’s responsibility to validate the performance of such product combination for any particular application.

A human A/T SNP in the AHRR7 gene was analyzed using genomic DNA from wild-type (blue), homozygous mutant (green), and heterozygous (red) samples. Experiments were performed using the Type-it HRM PCR Kit and a Rotor-Gene Q cycler with an HRM channel. Data analysis was performed with the unsupervised mode of Rotor-Gene ScreenClust HRM Software. A/T polymorphisms (class IV SNPs) are most difficult to discriminate due to minute differences between homozygote alleles (in this example, less than 0.1°C). [A] HRM raw data, [B] cluster plot. All pseudo-unknowns were correctly clustered according to genotype.|Twofold dilutions of human genomic DNA from 30 ng (10,000 copies) to 0.06 ng (20 copies) were used as a template in real-time PCR. Five replicate reactions were run for each dilution using a self-designed TaqMan® assay for IL1R2 and the Rotor-Gene Probe PCR kit on the Rotor-Gene Q. The average difference in the CT values between all dilutions was 1.07 cycles. Human genomic DNA was used as template in 72 replicate real-time PCRs using a self-designed TaqMan® assay for BCL2 on the Rotor-Gene Q without ROX normalization. The average CT value was 24.94 with a standard deviation of only 0.05, equivalent to a CV of 0.2%.

|Mixtures of methylated and unmethylated DNA (EpiTect Control DNA) of varying ratios were used as template. A CpG island from the promoter region of the APC gene (adenomatosis polyposis coli) was amplified and the degree of methylation was determined by HRM methylation analysis on the Rotor-Gene Q using the EpiTect HRM PCR Kit. [A] A standard normalized melt curve and a [B] difference plot normalized to the 50% methylated sample are shown.|Heating/cooling is achieved by rapid airflow in the reaction chamber. Tubes spin past the excitation/detection optics every 150 milliseconds enabling high-speed data capture. Up to 6 separate LED light sources can be used in combination with 6 different detection filters and a highly sensitive photomultiplier detector.|The Rotor-Gene Q supports multiple formats to suit a range of needs. Choose between tubes or Rotor-Discs, which offer accelerated setup and high throughput. Change the format in seconds by simply switching the snap-fit metal rotor that holds your plasticware format of choice.|

The Rotor-Gene Q is the only real-time cycler currently capable of deciphering the most difficult class IV SNPs by HRM. Harness the power of HRM using dedicated QIAGEN HRM Kits for applications such as genotyping (see figure "Identification of a class IV SNP" for data from the Type-it HRM PCR Kit), quantitative methylation analysis (see figure "Highly sensitive detection of methylated DNA" for data from the EpiTect HRM PCR Kit), gene scanning, and sequence matching. The Type-it HRM PCR Kit reliably and accurately detects gene mutations and SNPs. The EpiTect HRM PCR Kit enables fast screening and accurate detection of changes in CpG methylation status of bisulfite converted DNA.

Principle

Unique rotary design for outstanding performance

The unique centrifugal rotary design of the Rotor-Gene Q makes it the most precise and versatile real-time PCR cycler currently available (see figure "Cross-section of the Rotor-Gene Q"). Each tube spins in a chamber of moving air, keeping all samples at precisely the same temperature during rapid thermal cycling. Detection is similarly uniform. When each tube aligns with the detection optics, the sample is illuminated and the fluorescent signal is rapidly collected from a single, short optical pathway. This thermal and optical uniformity results in sensitive, precise, and fast real-time PCR analysis (see figure "Precise real-time PCR analysis"). It also eliminates sample-to-sample variations and edge effects. These are unavoidable in traditional block-based instruments due to temperature gradients across the block and multiple, complex optical pathways.

The rotary design delivers:

Tube-to-tube variation ±0.02°C

Uniform detection eliminating the need for ROX reference dye

Fast ramping and negligible equilibration times for short run-times

Complete confidence in your results

Unrivaled optical range enables multiple applications

Whether your assay is based on intercalating dyes such as SYBR® Green, probes such as hydrolysis (TaqMan®), hybridization (FRET), Scorpion probes, or other multiplex chemistries, the Rotor-Gene Q meets your requirements. With up to 6 channels spanning UV to infrared wavelengths, the cycler delivers the widest optical range currently available (see table). In addition, the software allows you to create new excitation/detection wavelength combinations, which means that the Rotor-Gene Q is compatible with dyes you may use in the future.

Channels for optical detection

Channel

Excitation (nm)

Detection (nm)

Examples of fluorophores detected

Blue

365 ± 20

460 ± 20

Marina Blue, Edans, Bothell Blue,
Alexa Fluor 350, AMCA-X

Green

470 ± 10

510 ± 5

FAM, SYBR® Green I, Fluorescein,
EvaGreen, Alexa Fluor 488

Yellow

530 ± 5

557 ± 5

JOE, VIC, HEX, TET, MAX, CAL Fluor Gold 540, Yakima Yellow

Orange

585 ± 5

610 ± 5

ROX, CAL Fluor Red 610, Cy 3.5,
Texas Red, Alexa Fluor 568

Red

625 ± 5

660 ± 10

Cy5, Quasar 670, LightCycler
Red640, Alexa Fluor 633

Crimson

680 ± 5

712 high pass

Quasar 705, LightCycler Red705,
Alexa Fluor 680

HRM

460 ± 20

510 ± 5

SYBR® Green I, SYTO9, LC Green,
LC Green Plus+, EvaGreen

Expand your research with HRM

High-resolution melting analysis (HRM) is a closed-tube, post-PCR analysis that has raised enormous scientific interest. HRM characterizes double-stranded PCR products based on their dissociation (melting) behavior. It is similar to classical melting curve analysis, but provides far more information for a wider range of applications. PCR products can be discriminated according to sequence, length, GC content, or strand complementarity, down to single base-pair changes. Previously unknown and even complex sequence variations can be readily detected and characterized in a robust and straightforward way. The rotary design of the Rotor-Gene Q and its outstanding thermal and optical performance are highly suited to HRM.

The HRM option for the Rotor-Gene Q includes:

A specially tuned high-intensity optical HRM channel

Thermal resolution of 0.02°C

High data-acquisition rates

Comprehensive HRM software

Minimum maintenance; maximum convenience

The Rotor-Gene Q is engineered to reduce the need for maintenance and to maximize ease of use. This saves time and costs, and allows you to focus on your research; not on keeping the cycler up and running.

Convenient features of the Rotor-Gene Q include:

Lifetime guarantee on highly stable LEDs, no expensive lamps or lasers to change, no gradual performance loss of light source

No optical calibration needed at installation or when the instrument is moved

No sample block to clean

No condensation or bubbles in reactions due to rotation

Small, light, and robust; simply place the instrument wherever you like

Procedure

Flexible formats match your workflows

The Rotor-Gene Q supports multiple PCR tube formats to suit a range of needs (see figure "Flexible formats"). Changing the format, by simply switching the snap-fit metal rotor that holds the tubes, takes just seconds. As well as tubes, Rotor-Discs are available, which offer accelerated setup and higher throughput. Rotor-Discs are circular plates of vertically oriented reaction wells. The Rotor-Disc 100 is the equivalent of a 96-well plate with an additional 4 reference wells. These extra wells can be conveniently used for more reactions or additional controls. Alternatively, the Rotor-Disc 72 has 72 wells. Rotor-Discs can be quickly and easily sealed with plastic film using a Rotor-Disc Heat Sealer. For all you need to run reactions using Rotor-Discs, choose the Rotor-Disc 100 Starter Kit or the Rotor-Disc 72 Starter Kit.

You can perform manual reaction setup, or take advantage of QIAGEN's automated solutions for reaction setup. The QIAgility is cost-effective and delivers rapid, high-precision PCR setup, while the QIAsymphony AS is ideal for laboratories performing routine PCR tests on a day-to-day basis. Both instruments perform automated reaction setup in Rotor-Gene formats, allow direct transfer of sample lists, and are supplied with verified protocols for real-time PCR master mixes.

Easy routine verification

Laboratories may often want to verify thermal accuracy. For most cyclers, this requires interaction with a service engineer. With the Rotor-Gene Q, this is not necessary. Instead, the easy-to-use, cost-effective Rotor-Disc OTV (Optical Temperature Verification) Kit automates accuracy testing. The full procedure takes only a couple of minutes.

Applications

A range of QIAGEN kits for the Rotor-Gene Q enables reliable quantification in all your real-time PCR applications without the need for optimization of reaction and cycling conditions. Kits for real-time PCR and HRM applications are available for:

Gene expression analysis

Pathogen detection

DNA methylation analysis

Genotyping and gene scanning

miRNA research

Software

Q-Rex Software

Q-Rex Software is a new operating and analysis software for the Rotor-Gene Q, providing several unique new features that promote a more user-friendly interface to help streamline your qPCR workflow. The software is suitable for use by the most novice researchers, while maintaining the highly complex data analysis functions required by advanced researchers.

Rotor-Gene Q Software

The comprehensive Rotor-Gene Q software package supports all current state-of-the art real-time analysis procedures from basic to advanced algorithms. This provides complete freedom to analyze your valuable experimental data and increases the reliability of your results. Data security is assured and all process steps are trackable from starting the run to exporting the results.

Superior software available for genotyping and mutation detection using HRM analysis

Rotor-Gene ScreenClust HRM Software is an extension to the Rotor-Gene operating software. This software is the most powerful tool currently available for analysis of HRM data from the Rotor-Gene Q or Rotor-Gene 6000 cycler. By grouping samples into clusters, Rotor-Gene ScreenClust HRM Software opens a new dimension in HRM analysis for applications such as genotyping and mutation screening.

REST software 2009

REST software 2009 is a standalone software tool for analysis of gene expression data from quantitative real-time PCR experiments. REST software 2009 is available for download under the "Resources" tab, and provides valuable analysis, including:

Estimation of up- and down-regulation for gene expression studies

Randomization and bootstrapping techniques

Graphical data output via whisker-box plots

Traditional relative quantification enables estimation of gene expression. However, this method does not provide statistical information that is suitable for comparing expression in groups of treated and untreated samples in a robust manner. The integrated randomization and bootstrapping methods used in REST software 2009 test the statistical significance of calculated expression ratios and can be used even when the data includes outliers.
REST software 2009 applies a mathematic model that takes into account the different PCR efficiencies of the gene of interest and reference genes. Compared to using a single reference gene, using multiple reference genes for normalization can improve the reliability of results.

For use on the Rotor-Gene Q. Rotor-Gene Q Software 2.3 is compatible with both Windows XP and Windows 7 operating systems. If you would like to upgrade the operating system for your QIAGEN computer (HP ProBook 6550b or HP ProBook 6570b) to Windows 7, please contact QIAGEN Technical Services for your Windows 7 image CD.

For use with Q-Rex Software version 1.0 to calculate absolute concentration of targets by PCR. This user manual provides information about the functions and features of the Q-Rex Absolute Quantification
HID plug-in.

For use with the Q-Rex Software v1.0 for determination of genotypes
using High-Resolution Melting analysis. This user manual provides information about the functions and features of the Q-Rex HRM Analysis plug-in.

For use with the Q-Rex Software v1.0 for determination of genotypes using Melt Curve Analysis. This user manual provides information about the functions and features of the Q-Rex Melt Curve Analysis
plug-in.